Jiménez-Pearson María-Antonieta, Delany Isabel, Scarlato Vincenzo, Beier Dagmar
Theodor-Boveri-Institut für Biowissenschaften, Lehrstuhl für Mikrobiologie, Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Molecular Immunology Unit, Chiron Vaccines, Via Fiorentina 1, 53100 Siena, Italy.
Microbiology (Reading). 2005 Oct;151(Pt 10):3299-3311. doi: 10.1099/mic.0.28217-0.
It is well established that motility is an essential virulence trait of the human gastric pathogen Helicobacter pylori. Accordingly, chemotaxis contributes to the ability of H. pylori to colonize animal infection models. Chemotactic signal transduction in H. pylori differs from the enterobacterial paradigm in several respects. In addition to a separate CheY response regulator protein (CheY1), H. pylori contains a CheY-like receiver domain (CheY2) which is C-terminally fused to the histidine kinase CheA. Furthermore, the genome of H. pylori encodes three CheV proteins consisting of an N-terminal CheW-like domain and a C-terminal receiver domain, while there are no orthologues of the chemotaxis genes cheB, cheR and cheZ. To obtain insight into the mechanisms controlling the chemotactic response of H. pylori, we investigated the phosphotransfer reactions between the purified two-component signalling modules in vitro. We demonstrate that both CheY1 and CheY2 are phosphorylated by CheA approximately P and that the three CheV proteins mediate the dephosphorylation of CheA approximately P, but with a clearly reduced efficiency as compared to CheY1 and CheY2. Furthermore, our data indicate retrophosphorylation of CheAY2 by CheY1 approximately P, suggesting a role of CheY2 as a phosphate sink to modulate the half-life of CheY1 approximately P.
众所周知,运动性是人类胃部病原体幽门螺杆菌的一种重要毒力特性。因此,趋化作用有助于幽门螺杆菌在动物感染模型中定殖。幽门螺杆菌中的趋化信号转导在几个方面不同于肠道细菌模式。除了一个单独的CheY应答调节蛋白(CheY1)外,幽门螺杆菌还含有一个CheY样受体结构域(CheY2),该结构域在C末端与组氨酸激酶CheA融合。此外,幽门螺杆菌的基因组编码三种CheV蛋白,它们由一个N末端CheW样结构域和一个C末端受体结构域组成,而趋化基因cheB、cheR和cheZ没有直系同源物。为了深入了解控制幽门螺杆菌趋化反应的机制,我们在体外研究了纯化的双组分信号模块之间的磷酸转移反应。我们证明,CheY1和CheY2都被CheA磷酸化形成CheAP,并且三种CheV蛋白介导CheAP的去磷酸化,但与CheY1和CheY2相比效率明显降低。此外,我们的数据表明CheY1P对CheAY2进行逆向磷酸化,这表明CheY2作为一个磷酸汇来调节CheY1P的半衰期。